FLUOROPATCH

Direct observation of ligand binding and channel gating on individual ligand-gated ion channels

 Coordinatore Universitätsklinikum Jena 

 Organization address city: Jena
postcode: 7740

contact info
Titolo: Ms.
Nome: Nicole
Cognome: Baier
Email: send email
Telefono: +49 3641 934350
Fax: +49 3641 933202

 Nazionalità Coordinatore Germany [DE]
 Totale costo 45˙000 €
 EC contributo 45˙000 €
 Programma FP7-PEOPLE
Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013)
 Code Call FP7-PEOPLE-2009-RG
 Funding Scheme MC-ERG
 Anno di inizio 2010
 Periodo (anno-mese-giorno) 2010-04-01   -   2013-06-30

 Partecipanti

# participant  country  role  EC contrib. [€] 
1    Nome Ente NON disponibile

 Organization address city: Jena
postcode: 7740

contact info
Titolo: Ms.
Nome: Nicole
Cognome: Baier
Email: send email
Telefono: +49 3641 934350
Fax: +49 3641 933202

DE (Jena) coordinator 45˙000.00

Mappa


 Word cloud

Esplora la "nuvola delle parole (Word Cloud) per avere un'idea di massima del progetto.

gated    clamp    directly    combine    time    ligand    binding    detection    molecular    confocal    accessible    channel    channels    patch    kinetics    fluorescence    alone    diffusion    gating    molecule    single    models   

 Obiettivo del progetto (Objective)

'Understanding the dynamics of ligand-gated ion channels is essential for understanding synaptic processes and neuronal signal integration (neurotransmitter gated channels) and intracellular signalling (cyclic nucleotide gated channels). Here we will combine single molecule fluorescence detection, with single channel electrophysiology, to detect ligand-binding and follow subsequent channel gating simultaneously. Single-channel patch-clamp alone can only measure two experimental states and their development in time: open and closed. Still, given some assumption, such data permitted construction of detailed kinetic models. Fluorescence detection can determines whether ligands are present or not, and the kinetics of the binding process. However, the nature of the ligand (agonist or antagonist) is not directly accessible. It was suggested more than 10 years [Edelstein 1997], that combining the two approaches on the single-molecule level will gain insights into the receptor mechanism. We will combine the two approaches to directly link the two molecular functional determining events (ligand binding and channel gating) within one experiment. Recently we implemented a combination of confocal single molecule fluorescence detection with single channel patch clamp, and showed (on the example of nAChR and fluorescent epibatidine) that such experiments are technically possible and feasible [manuscript in preparation]. We identified two parameters as limiting for the achievable time resolutions: counting noise and ligand diffusion into the confocal volume. Scope of this project includes: (1) identifying and analyzing biological systems with kinetics accessible to this approach, (2) developing techniques to reduce the time limitation, in particular the use of FRET to avoid the diffusion delay and (3) propose a model to describe the intra-molecular single transduction, and discuss it in comparison to models derived from single channel patch clamp alone.'

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